Structure of GDAP1 bound to a product of lipid peroxidation

与脂质过氧化产物结合的 GDAP1 的结构

基本信息

批准号：
10645396
负责人：
ANDREW Paul VANDEMARK
金额：
$ 7.23万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10645396
关键词：
Acceleration Active Sites Affect Affinity Animal Model Antioxidants Binding Binding Sites Biochemical Biochemistry Biological Biological Assay Brain Catalysis Cell model Cell physiology Cells Charcot-Marie-Tooth Disease Complex Cytoprotection Data Disease model Docking Drug Metabolic Detoxication Enzymatic Biochemistry Enzymes Event Foundations Future GDAP protein Genes Genetic Transcription Glutathione Glutathione S Transferase A Glutathione S-Transferase Goals Housing Inherited Investigation Ligand Binding Lipid Peroxidation Lipid Peroxides Lipids Location Malondialdehyde Measures Mediating Mitochondria Modeling Molecular Molecular Conformation Mutagenesis Mutation Neurons Neuropathy Outer Mitochondrial Membrane Oxidation-Reduction Oxidative Stress Peripheral Nervous System Diseases Persons Phenotype Play Poison Positioning Attribute Predisposition Proteins Resolution Role Shapes Signal Transduction Site Structure System Therapeutic Therapeutic Intervention Time Tooth Diseases United States Visualization X-Ray Crystallography Xenobiotics adduct autosome biological adaptation to stress interest knock-down member metabolic rate mitochondrial dysfunction molecular modeling molecular pathology mutant nervous system disorder overexpression oxidative damage protein protein interaction response small molecule success tool

项目摘要

Summary. Mutations in GDAP1 are associated with the peripheral neuropathy Charcot-Marie-Tooth disease. Charcot-Ma- rie-Tooth is one of most common inherited neurological disorders, estimated to affect 126,000 people in the United States alone. GDAP1 knockdown, overexpression, and as well as multiple models of CMT show changes consistent with dysregulation of the cellular response to oxidative stress. These include changes in NRF2-driven transcriptional activity, evidence of glutathione depletion, redox disbalance, and mitochondrial depolarization. At the same time key aspects of the mitochondrial network’s response to oxidative stress are very similar to key aspects of CMT phenotypes: fragmentation, fusion deficits and change in position inside the cell. Finally, GDAP1 is a member of the Glutathione S-transferase (GST) superfamily which protect the cell against peroxidated lipids and xenobiotics, toxic molecules that accumulate under conditions of oxidative stress. The mechanism underly- ing GDAP1’s role in oxidative stress response is unknown. We have recently discovered that GDAP1 can bind 4-hydroxynoneal (4HNE), a toxic lipid that is formed from the breakdown of peroxidated lipids primarily in the mitochondria. This proposal will address two main questions: can GDAP1 utilize 4HNE as a substrate, in a manner similar to canonical GST enzymes, or does it have a non-enzymatic role in the oxidative stress re- sponse? Secondly, are there conformational changes associated with or a consequence of 4HNE binding? We will address these questions by 1) biochemically measuring enzymatic parameters associated with 4HNE medi- ated GST activity; 2) biochemically defining the impact of important residues within the putative active site pocket on 4HNE binding affinity and GST enzymatic activity; 3) determining the structure of the GDAP1-4HNE complex using x-ray crystallography. These data will define the molecular mechanism by which GDAP1 recognizes 4HNE to facilitate binding and reveal and conformational changes in protein that are associated with 4HNE binding. If GDAP1 is an enzyme it will reveal how it facilitates catalysis of 4HNE with glutathione. If GDAP1 is playing a non-enzymatic role, conformational changes resulting from 4HNE will play a key role in GDAP1 function and will be revealed in this structure. Overall, these studies will be critical in shaping future biochemical and cell-based investigations centered on GDAP1 function. Further, the structure will provide the foundation needed to compu- tationally predict small molecule binding partners, critical tools for modulating GDAP1 function that will allow deeper interrogation of CMT disease models and a first step towards therapeutic intervention.

概括。 GDAP1中的突变与周围神经病型肉体 - 马里齿病有关。 charcot-ma- Rie Tooth是最常见的神经系统疾病之一，估计会影响126,000人仅美国。 GDAP1敲低，过表达以及CMT的多个模型显示变化与细胞对氧化应激反应的失调一致。这些包括变更NRF2驱动转录活性，谷胱甘肽耗竭的证据，氧化还原破坏和线粒体去极化。线粒体网络对氧化应激的响应的同一时间与关键非常相似 CMT表型的各个方面：碎片化，融合定义和变化细胞内部的位置。最后，GDAP1 是谷胱甘肽S-转移酶（GST）的成员，可保护细胞免受过氧化脂质的影响和异生物生物，在氧化应激条件下积累的有毒分子。基础机制 ING GDAP1在氧化应激反应中的作用尚不清楚。我们最近发现GDAP1可以绑定 4-羟基诺（4HNE），这是一种由过氧化脂质脂质的分解形成的有毒脂质线粒体。该建议将解决两个主要问题：GDAP1可以在类似于规范GST酶的方式，还是在氧化应激中具有非酶的作用赞助？其次，是否存在与4HNE结合相关的构象变化？我们将通过1）测量与4HNE MEDI-相关的酶参数来解决这些问题 ATED GST活动； 2）从生化来定义重要残留物在推定的活动现场袋中的影响在4HNE结合亲和力和GST酶活性上； 3）确定GDAP1-4HNE复合物的结构使用X射线晶体学。这些数据将定义GDAP1识别4HNE的分子机制促进与4HNE结合相关的蛋白质的结合，揭示和构象变化。如果 GDAP1是一种酶，它将揭示其如何用谷胱甘肽促进4HNE的催化。如果GDAP1在玩非酶作用，由4HNE产生的构象变化将在GDAP1函数中起关键作用，并且将在这个结构中揭示。总体而言，这些研究对于塑造未来的生化和基于细胞的研究至关重要以GDAP1功能为中心的调查。此外，该结构将为组成所需的基础预测小分子结合伙伴，调节GDAP1功能的关键工具，这将允许对CMT疾病模型的更深入询问和治疗干预的第一步。